AKT1/2/3 Antibody (YA633)

AKT1/2/3 Antibody (YA633) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to AKT1/2/3.

AKT1 Protein, a member of the AKT kinase family alongside AKT2 and AKT3, plays a pivotal role in regulating diverse cellular processes such as metabolism, proliferation, cell survival, growth, and angiogenesis. Functioning through serine and/or threonine phosphorylation of numerous downstream substrates, AKT1 is implicated in the modulation of over 100 reported substrate candidates, demonstrating its multifaceted nature. Notably, it orchestrates the insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter, impacting glucose uptake. Furthermore, AKT1 governs glucose storage as glycogen by phosphorylating GSK3A and GSK3B, inhibiting their kinase activity. Its involvement in various pathways includes mediating insulin-stimulated protein synthesis through the mTORC1 signaling pathway and regulating the FOXO factors, impacting cellular localization. AKT1 also influences NF-kappa-B-dependent gene transcription and positively regulates CREB1 activity, contributing to the transcription of pro-survival genes. Additionally, AKT1 exhibits a role in lipid metabolism by phosphorylating ATP citrate lyase (ACLY) and activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B), resulting in reduced cyclic AMP levels and inhibition of lipolysis. Beyond these functions, AKT1 is implicated in cell migration, adhesion assembly and disassembly, placental development, and diverse cellular responses to growth factors, including platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin, and insulin-like growth factor I (IGF-I). The recent discovery of its role in the SPATA13-mediated regulation of cell migration and adhesion adds to the complexity of its functions, highlighting AKT1's central role in cellular homeostasis and responsiveness to environmental cues.

Free

Q9Y243

Predicted band size: 56 kDa; Observed band size: 56 kDa

Protein A affinity purified.

Cell membrane, Cytoplasm, Membrane, Nucleus.

Non-conjugated

Unmodified

AB_3102049

Signal Transduction

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500-1:5,000; ICC/IF: 1:50-1:200; IHC-P: 1:50-1:200; FC: 1:50-1:100; IP: Use at an assay dependent concentration.

• 
  Western blot analysis of extracts from C6 (lane 2(20μg) and C6 (lane 3(40μg)using AKT1/2/3 (HY-P80009) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% defatted milk powder in TBST for 2 hour at room temperature. The primary antibody (HY-P80009, 1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% defatted milk powder in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of MCF-7 cells labeling AKT1/2/3 with AKT1/2/3 Antibody (YA633) (HY-P80009)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with AKT1/2/3 Antibody (YA633) (HY-P80009) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of RAW264.7cells labeling AKT1/2/3 with AKT1/2/3 Antibody (YA633) (HY-P80009) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with AKT1/2/3 Antibody (YA633) (HY-P80009)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using AKT1/2/3 Antibody (YA633). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80009, 1/2000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using AKT1/2/3 Antibody (YA633). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80009, 1/2000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, ICC/IF, IHC-P, IP, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human AKT3.AA range:300-479.